等位基因3'末端碱基特异性引物定量PCR检测人脂联素基因单核苷酸多态性

基础医学与临床 ›› 2007, Vol. 27 ›› Issue (9): 1025-1028.

等位基因3'末端碱基特异性引物定量PCR检测人脂联素基因单核苷酸多态性

宋现让王淑芳迟伟玲

山东省肿瘤医院基础研究中心济南市中心医院山东大学医学院

收稿日期:2005-06-14 修回日期:2006-10-08 出版日期:2007-09-25 发布日期:2007-09-25
通讯作者: 宋现让

Single Nucleotide Polymorphism of Human APM1 Detected by Using Allelic 3'-Terminal Specific Primers Real-time PCR

Xian-rang Song Shu-fang Wang Wei-ling Chi

Received:2005-06-14 Revised:2006-10-08 Online:2007-09-25 Published:2007-09-25
Contact: Xian-rang Song

摘要/Abstract

摘要： 目的建立非探针标记荧光定量PCR的SNP检测技术，分析人脂联素（APM1）基因单核苷酸多态性（SNP）与Ⅱ型糖尿病（T2DM）的关系。方法设计3'末端碱基特异性引物，进行定量PCR反应，通过比较各对引物的扩增效率判断基因型，并进行测序验证。利用该方法确定20例T2DM、24例肥胖及28例同年龄组正常人APM1基因的45位和276位碱基的SNP。结果该方法能有效地区分不同的基因型，与测序结果符合率100%。APM1基因45位SNP与T2DM相关（p<0.05），基因型为TT纯合子者发生T2DM的危险性是G/T和GG者的3倍；而肥胖与APM1基因45和276位点SNP无关。结论等位基因3'末端碱基特异性引物定量PCR可以用于基因SNP检测。APM基因45位碱基的SNP与T2DM发生有关。

Abstract: Objectives To develop an analyzing technique of single nucleotide polymorphism(SNP) based on non-probe real-time quantitative PCR, and to study the relationship between SNP in human adiponectin (APM1) gene and type 2 diabetes (T2DM). Methods Design primers with specific 3'-terminal nucleotides and perform Real-time PCR using SYBR Green dye. Genotypes were determined by comparing the amplification efficiency of each pair of primer and verified by sequencing. SNPs 45 and 276 in AMP of 20 T2DM, 24 obesity and 28 healthy persons were analyzed with this technique. Results This technique could discriminate genotype as efficient as sequencing. The genotype of APM1 at +45 site was significantly correlated with T2DM (p<0.05). Moreover, the relative risk of T2DM incidence in persons with TT genotype was 3 times of those with GG and GT. On the contrary, obesity was not correlated with SNP at +45 nor +276 sites of APM1 gene. Conclusion Allelic 3'-terminal specific primers real-time PCR could be used to determine SNPs. SNP at +45 sites of APM1 gene correlated with T2DM.

宋现让王淑芳迟伟玲. 等位基因3'末端碱基特异性引物定量PCR检测人脂联素基因单核苷酸多态性[J]. 基础医学与临床, 2007, 27(9): 1025-1028.

Xian-rang Song Shu-fang Wang Wei-ling Chi. Single Nucleotide Polymorphism of Human APM1 Detected by Using Allelic 3'-Terminal Specific Primers Real-time PCR[J]. Basic & Clinical Medicine, 2007, 27(9): 1025-1028.